EFFECTS OF NICOTINE ON AN IN VITRO RECONSTITUTED MODEL ORAL MUCOSA IN TERMS OF CYTOKINE PRODUCTION
AbstractBackground: The extensive use of tobacco and its associated problematic health issues have been aconcern to mankind. The World Health Organization (WHO) estimates that approximately one-third ofthe global population aged 15 years or older are smokers and each smoker consumes an average of 15cigarettes daily. The objective of this study was to establish the effect of nicotine on an in vitroreconstituted oral mucosa model, the effect of treatment with this compound was measured in terms ofcytokine production. Method: Observational laboratory based study design was used to carry out theexperiment. The reconstituted human epithelium model used in the study was prepared and supplied bySkin Ethic Laboratories, Nice, France. The effect of nicotine on epithelial cytokine production wasassessed using commercially available assay kits (R&D systems). This was done using the enzymelinked immuno-sorbent assay. Result: In this study there was evidence that after 5 minutes treatment onun-inflamed mucosa with nicotine at 10 mm concentration GM-CSF release decreased, and also after 24hours treatment with nicotine at 10mM concentration GM-CSF release increased. TNF-α increasedrelease of pro-inflammatory cytokines, IL-6, IL-8, and also GM-CSF from the model mucosa after 24hours, but had no effect on the release of IL-1α., IL-6, IL-8, and GM-CSF after 5 minutes and 24 hoursrespectively. Conclusion: In conclusion at all the concentrations used in this experiment, nicotine had noeffect on the TNF-α stimulated tissue and un-inflamed mucosa and had no significant effect on cytokinerelease including IL-1α., IL-6, IL-8, and GM-CSF after 5 minutes and 24 hours respectively.Keywords: Nicotine, oral mucosa, cytokine
WHO, Tobacco or health: a global status report. Geneva: World
Health Organization 1991 p. 1–48.
Mohamed S Al Moamary. Tobacco consummation: Is it still a
dilemma? Ann Thorac Med 2010;5(4):193–4.
Mannino DM. Chronic obstructive pulmonary disease: definition
and epidemiology. Respir Care 2003;48:1185–91, discussion
Tsiara S, Elisaf M, Mikhailidis DP. Influence of smoking on
predictors of vascular disease. Angiology 2003;54:507–30.
Tanski SE, Prokhorov AV, Klein JD. Youth and tobacco. Minerva
Lu GH, Ralapati S. Application of high-performance capillary
electrophoresis to the quantitative analysis of nicotine and
profiling of other alkaloids in ATF-regulated tobacco products.
Baldwin IT. Mechanism of damage-induced alkaloid production
in wild tobacco. J Chem Ecology 1989;15:1661–80.
Hashimoto T, Yamada Y. Alkaloid biogenesis: molecular aspects.
Annual Review of Plant Physiology and Plant Molecular Biology
Taybos G. Oral changes associated with tobacco use. Am J Med
Sham AS, Cheung LK, Jin LJ, Corbet EF. The effects of tobacco
use on oral health. Hong Kong Med J 2003;9:271–7.
Mirbod SM, Ahing SI. Tobacco-associated lesions of the oral
cavity: Part I. Nonmalignant lesions. J Can Dent Assoc
Dinarello CA. Interleukin-1, interleukin-1 receptors and interleukin-
receptor antagonist. Int Rev Immunol 1998;16:457–99.
Kupper TS, Min K, Sehgal P, Mizutani H, Birchall N, Ray A,
May L. Production of IL-6 by keratinocytes. Implications for
epidermal inflammation and immunity. Ann N Y Acad Sci
;557:454–64, discussion 464–5.
Taub DD, Anver M, Oppenheim JJ, Longo DL, Murphy WJ. T
lymphocyte recruitment by interleukin-8 (IL-8). IL-8-induced
degranulation of neutrophils releases potent chemoattractants for
human T lymphocytes both in vitro and in vivo. J Clin Invest
Li J, Ireland GW, Farthing PM, Thornhill MH. Epidermal and oral
keratinocytes are induced to produce RANTES and IL-8 by
cytokine stimulation. J Invest Dermatol 1996;106:661–6.
Braunstein S, Kaplan G, Gottlieb AB, Schwartz M, Walsh G,
Abalos RM, et al. GM-CSF activates regenerative epidermal
growth and stimulates keratinocyte proliferation in human skin in
vivo. J Invest Dermatol 1994;103:601–4.
Lee JH, Kim NS, Kwon TH, Jang YS, Yang MS. Increased
production of human granulocyte-macrophage colony stimulating
factor (hGM-CSF) by the addition of stabilizing polymer in plant
suspension cultures. J Biotechnol 2002;96(3):205–11.
Klapproth H, Racké K, Wessler I. Acetylcholine and nicotine
stimulate the release of granulocyte-macrophage colony
stimulating factor from cultured human bronchial epithelial cells.
Naunyn Schmiedebergs Arch Pharmacol 1998;357:472–5.
Rioux N, Castonguay A. 4-(methylnitrosamino)-1-(3-pyridyl)-1-
butanone modulation of cytokine release in U937 human
macrophages. Cancer Immunol Immunother 2001;49:663–70.
Kwon OS, Chung JH, Cho KH. Nicotine enhanced epithelial
differentiation in reconstructed human oral mucosa in vitro. Skin
Pharmacol Appl Skin Physio 1999;12:227–34.
Hoffmann D, Hoffmann I. The changing cigarette, 1950-1995. J
Toxicol Environ Health 1997;50(4):307–64.
Hecht SS. Biochemistry, biology, and carcinogenicity of tobaccospecific N-nitrosamines. Chem Res Toxicol 1998;11:559–603.
Squier CA, Johnson GK. Role of nicotine as a cofactor in
smokeless tobacco carcinogenesis. Smoking and Tobacco Control
Monograph 2. NIH Pub 92;1992.p. 153–60.
Dussor GO, Leong AS, Gracia NB, Kilo S, Price TJ, Hargreaves
KM. Potentiation of evoked calcitonin gene- related peptide release
from oral mucosa: a potential basis for the pro- inflammatory
effects of nicotine. Eur J Neuro Sci 2003;18:2515–26.
Barker JN, Mitra RS, Griffiths CE, Dixit VM, Nickoloff BJ.
Keratinocytes as initiators of inflammation. Lancet
Johnson GK, Organ CC. Prostaglandin E2 and interleukin-1
concentrations in nicotine-exposed oral keratinocyte cultures. J
Periodontal Res 1997;32:447–54.
Rupniak HT, Rowlatt C, Lane EB, Steele JG, Trejdosiewicz LK,
Laskiewicz B, Povey S, Hill BT. Characteristics of four new
human cell lines derived from squamous cell carcinomas of the
head and neck. J Natl Cancer Inst 1985;75(4):621–35.
Rosdy M, Clauss LC. Terminal epidermal differentiation of
human keratinocytes grown in chemically defined medium on
inert filter substrates at the air-liquid interface. J Invest Dermatol
Christen AG. The case against smokeless tobacco: five facts for
the health professional to consider. J Am Dent Assoc
Yamamoto T, Osaki T, Yoneda K, Ueta E. Cytokine production
by keratinocytes and mononuclear infiltrates in oral lichen planus.
J Oral Pathol Med 1994;23:309–15.
Kupper TS, Lee F, Birchall N, Clark S, Dower S. Interleukin 1
binds to specific receptors on human keratinocytes and induces
granulocyte macrophage colony-stimulating factor mRNA and
protein. A potential autocrine role for interleukin 1 in epidermis. J
Clin Invest 1988;82:1787–92.
Partridge M, Chantry D, Turner M, Feldmann M. Production of
interleukin-1 and interleukin-6 by human keratinocytes and
squamous cell carcinoma cell lines. J Invest Dermatol
Cheng YA, Tsai CC. Nicotine- and arecoline-induced interleukin-
secretion and intercellular adhesion molecular-1 expression in
human oral epidermoid carcinoma cells in vitro. Arch Oral Biol
Nakano Y, Kobayashi W, Sugai S, Kimura H, Yagihashi S.
Expression of tumor necrosis factor-alpha and interleukin-6 in oral
squamous cell carcinoma. Jpn J Cancer Res 1999;90:858–66.
Hong SH, Ondrey FG, Avis IM, Chen Z, Loukinova E, Cavanaugh
PF Jr, et al. Cyclooxygenase regulates human oropharyngeal
carcinomas via the proinflammatory cytokine IL-6: a general role
for inflammation? FASEB J 2000;14(11):1499–507.
Wendell KJ, Stein SH. Regulation of cytokine production in
human gingival fibroblasts following treatment with nicotine and
lipopolysaccharide. J Periodontol 2001;72:1038–44.
Sugiyama A, Uehara A, Iki K, Matsushita K, Nakamura R,
Ogawa T, et al. Activation of human gingival epithelial cells by
cell-surface components of black-pigmented bacteria:
augmentation of production of interleukin-8, granulocyte colonystimulating factor and granulocyte-macrophage colonystimulating factor and expression of intercellular adhesion
molecule 1. J Med Microbiol 2002;51:27–33.
Gabri MR, Menna PL, Scursoni AM, Gomez DE, Alonso DF.
Role of tumor-derived granulocyte-macrophage colonystimulating factor in mice bearing a highly invasive and metastatic
mammary carcinoma. Pathobiology 1999;67:180–5.
Jablonska E, Piecuch J, Piotrowski L, Grabowska Z. GM-CSF in
the culture supernatants of neutrophils and serum levels in patients
with inflammation and squamous cell carcinoma of oral cavity.
Pol Merkuriusz Lek. 2001;11:394–7.